• Title, Summary, Keyword: Inside-out patch

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Effects of Adenosine on the Ionic Channel Activated by Metabolic Inhibition in Rabbit Ventricular Myocytes

  • Han, Jin;Kim, Eui-Yong;Ho, Won-Kyung;Earm, Yung-E
    • The Korean journal of physiology & pharmacology
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    • v.30 no.1
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    • pp.1-9
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    • 1996
  • The objective of the present study was to characterize the role of adenosine in regulation of ATP-sensitive $K^+\;channel\;(K_{ATP}\;channel)$ activity in isolated rabbit ventricular myocytes using the patch clamp technique. Internal adenosine had little effects on KaTr channel activity. In an outside-out patch with intrapipette GTP and ATP, external adenosine stimulated $K_{ATP}\;channel$ activity. In an inside-out Patch with intrapipette adenosine, ATP reduced $K_{ATP}\;channel$ activity, and GTP stimulated $K_{ATP}\;channel$ activity. Adenosine receptor activation shifted the half-maximal inhibition Of $K_{ATP}\;channel\;from\;70\;to\;241\;{\mu}m$. These results Suggest that activation of adenosine receptors stimulates $K_{ATP}\;channels$ in rabbit ventricular myocytes by reducing the apparent affinity of the channel for ATP. The effect may be important for activating $K_{ATP}\;channels$ during early phase of myocardial ischemia.

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Effect of Prostaglandins $D_2,\;E_2\;and\;I_2\;on\;the\;Regulation\;of\;K_{ATP}$ Channel Activity in Rat Cardiac Myocytes

  • Ju, Jeong-Min;Nah, Seung-Yeol;Kim, Jae-Ha
    • The Korean Journal of Physiology and Pharmacology
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    • v.3 no.5
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    • pp.507-512
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    • 1999
  • Contribution of prostaglandins $D_2,\;E_2\;and\;I_2\;(PGD_2,\;PGE_2\;and\;PGI_2)$ on the regulation of ATP-sensitive $K^+$ channel $(K_{ATP}\;channel)$ was investigated in isolated single rat ventricular cardiac myocytes using the patch clamp technique. $PGD_2,\;PGE_2\;and\; PGI_2$ did not affect $K_{ATP}$ channel activity in the inside-out patch, but increased channel activity in a dose-dependent manner when the channel activities were attenuated by the administration of 100 ${\mu}M$ ATP to the internal solution in the inside-out patch. Channel activations by the prostaglandins were abolished by 50 ${\mu}M$ glibenclamide, a $K_{ATP}$ channel blocker. Dose-response curves of relative channel activity against the ATP concentrations of internal solution in the inside-out patch were shifted to the right in the presence of those three prostaglandins. The rank order of the channel stimulatory potencies $(as\;IC_{50}\;for\;ATP)$ calculated from the dose-response curves were $PGI_2\;>\;PGD_2\;>\;PGE_2.$ Conductance of the channel was not changed by those three prostaglandins. In conclusion, we suggest that prostaglandins $D_2,\;E_2\;and\;I_2$ are involved in the regulation of $K_{ATP}$ channel activity in certain circumstances, and that those three prostaglandins may cause myocardial relaxation by opening $K_{ATP}$ channels, thus protecting the heart from ischema.

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Modulation of $Ca^{2+}-Activated$ Potassium Channels by cGMP-Dependent Signal Transduction Mechanism in Cerebral Arterial Smooth Muscle Cell of the Rabbit

  • Han, Jin;Kim, Na-Ri;Lee, Kwang-Bok;Kim, Eui-Yong
    • The Korean Journal of Physiology and Pharmacology
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    • v.4 no.6
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    • pp.445-453
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    • 2000
  • The present investigation tested the hypothesis that the activation of protein kinase G (PKG) leads to a phosphorylation of $Ca^{2+}-activated$ potassium channel $(K_{Ca}\;channel)$ and is involved in the activation of $K_{Ca}$ channel activity in cerebral arterial smooth muscle cells of the rabbit. Single-channel currents were recorded in cell-attached and inside-out patch configurations of patch-clamp techniques. Both molsidomine derivative 3-morpholinosydnonimine-N-ethylcarbamide $(SIN-1,\;50\;{\mu}M)$ and 8-(4-Chlorophenylthio)-guanosine-3',5'-cyclic monophosphate $(8-pCPT-cGMP,\;100\;{\mu}M),$ a membrane-permeable analogue of cGMP, increased the $K_{Ca}$ channel activity in the cell-attached patch configuration, and the effect was removed upon washout of the drugs. In inside-out patches, single-channel current amplitude was not changed by SIN-1 and 8-pCPT-cGMP. Application of ATP $(100\;{\mu}M),$ cGMP $(100\;{\mu}M),$ ATP+cGMP $(100\;{\mu}M\;each),$ PKG $(5\;U/{\mu}l),$ ATP $(100\;{\mu}M)+PKG\;(5\;U/{\mu}l),$ or cGMP $(100\;{\mu}M)+PKG\;(5\;U/{\mu}l)$ did not increase the channel activity. ATP $(100\;{\mu}M)+cGMP\;(100\;{\mu}M)+PKG\;(5\;U/{\mu}l)$ added directly to the intracellular phase of inside-out patches increased the channel activity with no changes in the conductance. The heat-inactivated PKG had no effect on the channel activity, and the effect of PKG was inhibited by 8-(4-Chlorophenylthio)-guanosine-3',5'-cyclic monophosphate, Rp-isomer $(Rp-pCPT-cGMP,\;100\;{\mu}M),$ a potent inhibitor of PKG or protein phosphatase 2A (PP2A, 1 U/ml). In the presence of okadaic acid (OA, 5 nM), PP2A had no effect on the channel activity. The $K_{Ca}$ channel activity spontaneously decayed to the control level upon washout of ATP, cGMP and PKG, and this was prevented by OA (5 nM) in the medium. These results suggest that the PKG-mediated phosphorylations of $K_{Ca}$ channels, or some associated proteins in the membrane patch increase the activity of the $K_{Ca}$ channel, and the activation may be associated with the vasodilating action.

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Design of a Ceramic Patch Antenna for Artillery Projectile Positioning Applications

  • Chae, Yoon-Kyoung;Lee, Chan-Ho;Cha, Jae-Ho;Kim, Wan-Joo;Ahn, Bierng-Chearl
    • 한국정보기술학회논문지
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    • v.15 no.5
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    • pp.81-85
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    • 2017
  • This paper presents a compact circularly-polarized patch antenna used for GPS/GLONASS receptions onboard an artillery projectile fuze. The proposed antenna consists of a corner-truncated slotted ceramic patch and a small circular ground plane. Two small rectangular slots are realized on the patch for improving the circular polarization bandwidth and an additional truncation at the patch's supper left corner is employed for enhancing the required impedance bandwidth. The designed antenna inside the fuze occupies a cylindrical space of 6-mm height and 36-mm diameter. The optimal antenna design and the analysis of effects on design parameters are carried out with Microwave Studio$^{TM}$ by CST. The fabricated antenna is to be installed inside a fuze radome potted with plastic foam. The fabricated antenna shows a gain of 2.5~4.2dBic, an axial ratio of 5.1~11.6dB, and a reflection coefficient of less than -10dB at 1.57~1.62GHz.

Activation of ATP-sensitive Potassium Channels by the Predominant Metabolite of Isoflurane in Rabbit Ventricular Myocytes

  • Han, Jin;Kim, Na-Ri;Kim, Eui-Yong;Kim, Sung-Ju;Cho, Kang-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.2
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    • pp.165-175
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    • 2001
  • Background: Recent in vivo experimental evidence suggests that isoflurane-induced cardioprotection may involve $K_{ATP}$ channel activation. However, it was demonstrated that isoflurane inhibited $K_{ATP}$ channel activities in the inside-out patch mode. To explain this discrepancy, the present investigation tested the hypothesis that a metabolite of isoflurane, trifluoroacetic acid (TFA), contributes to isoflurnae-induced cardioprotection via $K_{ATP}$ channel activation during myocardial ischemia and reperfusion. Methods: Single ventricular myocytes were isolated from rabbit hearts by an enzymatic dissociation procedure. Patch-clamp techniques were used to record single-channel currents. $K_{ATP}$ channel activities were assessed before and after the application of TFA with the inside-out patch mode. Results: TFA enhanced channel activity in a concentration-dependent fashion. The concentration of TFA for half-maximal activation and the Hill coefficient were 0.03 mM and 1.2, respectively. TFA did not affect the single channel conductance of $K_{ATP}$ channels. Analysis of open and closed time distributions showed that TFA increased burst duration and decreased the interburst interval without changes in open and closed time distributions shorter than 5 ms. TFA diminished ATP sensitivity of $K_{ATP}$ channels in a concentration-response relationship for ATP. Conclusions: TFA, a metabolite of isoflurane, enhanced $K_{ATP}$ channel activity in a concentration-dependent fashion. These results imply that TFA could mediate isoflurane-induced cardioprotection via $K_{ATP}$ channel activation during myocardial ischemia and reperfusion.

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Effects of $Cl^-$ Channel Blockers on the Cardiac ATP-sensitive $K^+$ Channel

  • Kwak, Yong-Geun
    • The Korean Journal of Physiology and Pharmacology
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    • v.3 no.3
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    • pp.305-313
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    • 1999
  • To explore whether $Cl^-$ channel blockers interact with the ATP-sensitive $K^+\;(K_{ATP})$ channel, I have examined the effect of two common $Cl^-$ channel blockers on the $K_{ATP}$ channel activity in isolated rat ventricular myocytes using patch clamp techniques. In inside-out patches, 4,4'-diisothio-cyanatostilbene- 2,2'-disulfonic acid (DIDS) and niflumic acid applied to bath solution inhibited the $K_{ATP}$ channel activity in a concentration-dependent manner with $IC_{50}$ of 0.24 and 927 ${\mu}M,$ respectively. The inhibitory action of DIDS was irreversible whereas that of niflumic acid was reversible. Furthermore, DIDS-induced block was not recovered despite exposure to ATP (1 mM). In cell-attached and inside-out patches, DIDS blocked the pinacidil- or 2,4-dinitrophenol (DNP)-induced $K_{ATP}$ channel openings. In contrast, niflumic acid did not block the pinacidil-induced $K_{ATP}$ channel openings in inside-out patches, but inhibited it in cell-attached patches. DIDS and niflumic acid produced additional block in the presence of ATP and did not affect current-voltage relationship and channel kinetics. All these results indicate that DIDS among $Cl^-$ channel blockers specifically blocks the cardiac $K_{ATP}$ channel.

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DIRECT MODULATI6ON OF MAXI-K CHANNEL IN SMOOTH MUSCLE CELL

  • Lee, Moo-Yeol;Chung, Sung-Kwon;Bang, Hyo-Weon;Uhm, Dae-Yong
    • Proceedings of the Korean Biophysical Society Conference
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    • pp.36-36
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    • 1996
  • The activities of Maxi-K channels were recorded using inside-out patches. The application of 30 nM of non-specific G protein activator, GTP $\gamma$S, to the intracellular side of the channels increases the channel activities about 3-fold, indicating that there exist some G proteins within the patch membranes to regulate the channel activities. (omitted)

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The Effects of Intracellular Monocarboxylates on the ATP-sensitive Potassium Channels in Rabbit Ventricular Myocytes

  • Kim, Na-Ri;Han, Jin;Kim, Eui-Yong;Ho, Won-Kyung;Earm, Yung E.
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.5
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    • pp.581-589
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    • 1998
  • A regulating mechanism of the ATP-sensitive potassium channels $(K_{ATP}\;channels)$ is yet to fully explained. This study was carried out to investigate the effects of intracellular application of monocarboxylates (acetate, formate, lactate, and pyruvate) on $K_{ATP}$ channels in isolated rabbit ventricular myocytes. Single channel currents of $K_{ATP}$ channels were recorded using the excised inside-out or permeabilized attached (open-cell) patch-clamp technique at room temperature. Intracellular application of acetate, formate and pyruvate led to an inhibition of channel activity, whereas intracellular application of lactate increased channel activity. These effects were reversible upon washout. Analysis of single channel kinetics showed that monocarboxylates did not affect open-time constant and close-time constant. These results suggest that monocarboxylates participate in modulating $K_{ATP}$ channels activity in cardiac cells and that modulation of $K_{ATP}$ channels activity may resolve the discrepancy between the low $K_i$ in excised membrane patches and high levels of intracellular ATP concentration during myocardial ischemia or hypoxia.

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Stretch-activated $K^+$ Channels in Rat Atrial Myocytes

  • Youm, Jae-Boum
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.6
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    • pp.341-348
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    • 2003
  • Mechanical stimuli to the cardiac myocytes initiate many biochemical and physiological events. Stretch-activated cation channels have been suggested to mediate these events. In this study, cell-attached and inside-out excised-patch clamp methods were used to identify stretch-activated cation channels in adult rat atrial myocytes. Channel openings were increased in cell-attached configuration when negative pressure was applied to the pipette, and also in inside-out excised patches by negative pressure. The channel was not permeable to $Cl^-$, $Na^+$ and $Cs^+$, but selectively permeable to $K^+$, and the degree of activation was dependent on the magnitude of negative pressure (full activation at ${\sim} -50 mmHg). In symmetrical 140 mM KCl, the slope conductance was $51.2{\pm}3$ pS between the potentials of -80 and 0 mV and $55{\pm}6$ pS between 0 and +80 mV (n=5). Glibenclamide ($100{mu}M$) or ATP (2 mM) failed to block the channel openings, indicating that it is not ATP-sensitive $K^+$ channel. Arachidonic acid ($30{mu}M$), which has been shown to activate a $K^+$ channel cooperatively with membrane stretch, did not affect the channel activity. $GdCl_3$ ($100{mu}M$) also did not alter the activity. These results demonstrate that the mechanical stretch in rat atrial myocytes activates a novel $K^+$-selective cation channel, which is not associated with other $K^+$ channels such as ATP-sensitive and arachidonic acid-activated $K^+$ channel.

Effect of pH on Calcium-Activated Potassium Channels in Pulmonary Arterial Smooth Muscle Cells of the Rabbit

  • Lee, Suk-Ho;Ho, Won-Kyung;Earm, Yung-E
    • The Korean journal of physiology & pharmacology
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    • v.25 no.1
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    • pp.17-26
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    • 1991
  • Single smooth muscle cells of the rabbit pulmonary artery were isolated by treatment with collagenase and elastase. Using the patch clamp technique, potassium channel activity was recorded from the inside-out membrane patch. The channel had a sin히e channel conductance of about 360 pS in symmetrical concentration of K on both sides of the patch, 150 mM, and had a linear current-voltage relationship. During the application of 10 mM tetraethylammonium (TEA) to the intracellular membrane surface, the amplitude of single channel current was reduced and very rapid flickering appeared. The open probability $(P_0)$ of this channel was increased by increasing positivity of the potential across the patch membrane, with e-fold increase by 20 mV depolarization, and by increasing the internal $Ca^{2+}$ concentration. These findings are consistent with those of large conductance Ca-activated K channels reported in other tissues. But the shortening of the mean open time by increasing $[Ca^{2+}]_i$, was an unexpected result and one additional closed state which might be arisen from a block of the open channel by Ca binding was suggested. The $P_0-membrane$ potential relationship was modulated by internal pH. Decreasing pH reduced $P_0$. Increasing pH not only increased $P_0$ but also weakened the voltage dependency of the channel opening. The modulation of Ca-activated K channel by pH was thought to be related to the mechanism of regulation of vascular tone by the pH change.

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